Application | Comment | Organism |
---|---|---|
pharmacology | conservation of the NTP-binding pocket among viruses of the family Flaviviridae as potential for development of therapeutics | Japanese encephalitis virus |
Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21 (DE3), recombinant protein, pET21b vector | Japanese encephalitis virus |
Crystallization (Comment) | Organism |
---|---|
enzymatically active fragment of the JEV NTPase/helicase catalytic domain, recombinant protein, crystal structure determined at 1.8 A resolution, data collection and refinement statistics | Japanese encephalitis virus |
Protein Variants | Comment | Organism |
---|---|---|
G199A | mutation in WALKER A motif, PCR-based mutagenesis, ATPase and RNA helicase activity lost | Japanese encephalitis virus |
G460A | mutation of residues of the arginine finger within the active sites of ATP hydrolysis, no effect on either ATPase or RNA-unwinding activities | Japanese encephalitis virus |
G463A | mutation of residues of the arginine finger within the active sites of ATP hydrolysis, no effect on either ATPase or RNA-unwinding activities | Japanese encephalitis virus |
K200A | mutation in WALKER A motif, PCR-based mutagenesis, ATPase and RNA helicase activity lost | Japanese encephalitis virus |
K200D | PCR-based mutagenesis, ATPase and RNA helicase activity lost | Japanese encephalitis virus |
K200E | PCR-based mutagenesis, ATPase and RNA helicase activity lost | Japanese encephalitis virus |
K200H | PCR-based mutagenesis, ATPase and RNA helicase activity lost | Japanese encephalitis virus |
K200N | PCR-based mutagenesis, ATPase and RNA helicase activity lost | Japanese encephalitis virus |
K200Q | PCR-based mutagenesis, ATPase and RNA helicase activity lost | Japanese encephalitis virus |
K200R | PCR-based mutagenesis, ATPase and RNA helicase activity lost | Japanese encephalitis virus |
Q457A | mutation of residues of the arginine finger within the active sites of ATP hydrolysis, 80% reduction of ATPase activity, no RNA helicase activity | Japanese encephalitis virus |
R458A | mutation of residues of the arginine finger within the active sites of ATP hydrolysis, 90% reduction of ATPase activity, no RNA helicase activity | Japanese encephalitis virus |
R459A | mutation of residues of the arginine finger within the active sites of ATP hydrolysis, no effect on either ATPase or RNA-unwinding activities | Japanese encephalitis virus |
R461A | mutation of residues of the arginine finger within the active sites of ATP hydrolysis, no ATPase activity, no RNA helicase activity | Japanese encephalitis virus |
R464A | mutation of residues of the arginine finger within the active sites of ATP hydrolysis, no ATPase activity, no RNA helicase activity | Japanese encephalitis virus |
T201A | mutation in WALKER A motif, PCR-based mutagenesis, ATPase and RNA helicase activity lost | Japanese encephalitis virus |
V462A | mutation of residues of the arginine finger within the active sites of ATP hydrolysis, no effect on either ATPase or RNA-unwinding activities | Japanese encephalitis virus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | no ATPase activity of the wild-type in the absence of | Japanese encephalitis virus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
54000 | - |
molecular mass of the helicase/NTPase domain, SDS-PAGE | Japanese encephalitis virus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + H2O | Japanese encephalitis virus | - |
ADP + phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Japanese encephalitis virus | P27395 | - |
- |
Purification (Comment) | Organism |
---|---|
gel filtration, recombinant protein | Japanese encephalitis virus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
structural characterization of catalytic domain, mutation analysis of residue substitution in the Walker A motif (Gly199, Lys200 and Thr201), within the NTP-binding pocket (Gln457, Arg461 and Arg464) and of Arg458 in the outside of the pocket in the motif IV, residues crucial for ATPase and RNA helicase activities and virus replication, Lys200 cannot be substituted by other residues to establish sufficient activities, structure of the NTP-binding pocket well conserved among the viruses of the Flaviviridae | Japanese encephalitis virus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + H2O | - |
Japanese encephalitis virus | ADP + phosphate | - |
? | |
ATP + H2O | genome structure, crystals and three-dimensional structure determined, structure of NTP-binding region, conserved residues within the NTP-binding pocket, ATPase and RNA helicase activities determined | Japanese encephalitis virus | ADP + phosphate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
nonstructural protein 3 | ambiguous | Japanese encephalitis virus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Japanese encephalitis virus |